Järhult Josef D, Wahlgren John, Hasan Badrul, Salaneck Erik, Lundkvist Åke
Section for Infectious Diseases, Department of Medical Sciences, Uppsala University, Uppsala, Sweden.
Zoonosis Science Center, Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala, Sweden;
Infect Ecol Epidemiol. 2015 Sep 8;5:28458. doi: 10.3402/iee.v5.28458. eCollection 2015.
To date, the most efficient and robust method for isolating avian influenza A viruses (IAVs) is using embryonated chicken eggs (ECEs). It is known that low-pathogenic avian IAVs undergo rapid genetic changes when introduced to poultry holdings, but the factors driving mutagenesis are not well understood. Despite this, there is limited data on the effects of the standard method of virus isolation of avian-derived viruses, that is, whether isolation in ECEs causes adaptive changes in avian IAVs. Eggs from a homologous species could potentially offer an isolation vessel less prone to induce adaptive changes.
We performed eight serial passages of two avian IAVs isolated from fecal samples of wild Mallards in both ECEs and embryonated Mallard eggs, and hemagglutination assay titers and hemagglutinin sequences were compared.
There was no obvious difference in titers between ECEs and embryonated Mallard eggs. Sequence analyses of the isolates showed no apparent difference in the rate of introduction of amino acid substitutions in the hemagglutinin gene (three substitutions in total in embryonated Mallard eggs and two substitutions in ECEs).
Embryonated Mallard eggs seem to be good isolation vessels for avian IAVs but carry some practical problems such as limited availability and short egg-laying season of Mallards. Our study finds isolation of Mallard-derived avian IAVs in ECEs non-inferior to isolation in embryonated Mallard eggs, but more research in the area may be warranted as this is a small-scale study.
迄今为止,分离甲型禽流感病毒(IAVs)最有效且可靠的方法是使用鸡胚(ECEs)。已知低致病性禽流感病毒引入家禽养殖场后会迅速发生基因变化,但驱动诱变的因素尚不清楚。尽管如此,关于禽源病毒标准病毒分离方法的影响的数据有限,即ECEs中的分离是否会导致禽流感病毒发生适应性变化。来自同源物种的鸡蛋可能提供一个不易诱导适应性变化的分离容器。
我们对从野生绿头鸭粪便样本中分离出的两种禽流感病毒在ECEs和绿头鸭胚蛋中进行了八次连续传代,并比较了血凝试验效价和血凝素序列。
ECEs和绿头鸭胚蛋之间的效价没有明显差异。分离株的序列分析表明,血凝素基因中氨基酸替换的引入率没有明显差异(绿头鸭胚蛋中总共三个替换,ECEs中两个替换)。
绿头鸭胚蛋似乎是禽流感病毒的良好分离容器,但存在一些实际问题,如绿头鸭的可获得性有限和产蛋季节短。我们的研究发现,在ECEs中分离绿头鸭源禽流感病毒并不比在绿头鸭胚蛋中分离差,但由于这是一项小规模研究,该领域可能需要更多研究。